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Function of KpsS, KpsC, and NeuE in Escherichia coli K1 Polysialic Acid Biosynthesis
Author(s) -
Thon Vireak,
Peterson Dwight,
Vionnet Justine,
Vann Willie
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.718.12
Subject(s) - polysialic acid , escherichia coli , biochemistry , biosynthesis , chemistry , function (biology) , gene , biology , genetics , cell adhesion , cell , neural cell adhesion molecule
Pathogenic Escherichia coli are often encapsulated with acidic polysaccharides such as polysialic acid. We have shown that four genes, kpsS , kpsC, neuE and neuS , in the E. coli K1 gene cluster are essential for initiation and elongation in E. coli K1 polysialic acid biosynthesis. NeuS is a polysialyltransferase. A previous report demonstrated that homologues of kpsC and kpsS gene products in N. meningitidis are cytidine 5′‐monophosphate‐2‐keto‐3‐deoxyoctulosonate (CMP‐KDO) transferases. These enzymes are involved in the biosynthesis of the putative glycolipid terminus of terminus of polysialic acid. Our objective in this report is to determine the function of KpsC, KpsS, and NeuE of E. coli in initiation of polysialic acid synthesis. We have successfully expressed in a soluble form and purified KpsC, KpsS, and NeuE of E. coli K1 to near homogeneity. This is the first time these full length membrane proteins have been expressed in a soluble form. We used HPLC and TLC methods to demonstrate that E. coli K1 KpsS and KpsC can transfer KDO to a fluorescence glycolipid acceptor substrate. The fluorescent products of KpsS and KpsC were isolated and used to investigate the function of the NeuE in polysialic acid biosynthesis.